Fig. 4: CircRNA hsa_circ_0007813 acted as a sponge for hsa-miR-361-3p, a micro-RNA targeting IGF2R. | Cell Death & Disease

Fig. 4: CircRNA hsa_circ_0007813 acted as a sponge for hsa-miR-361-3p, a micro-RNA targeting IGF2R.

From: Autophagy-associated circular RNA hsa_circ_0007813 modulates human bladder cancer progression via hsa-miR-361-3p/IGF2R regulation

Fig. 4

A Venn diagram showed potential downstream miRNA targets of hsa_circ_0007813. The prediction was carried out by interactome, circBank, and miRanda. B RNA-RNA pulldown assays were carried out to determine the downstream miRNA (N = 3, Kruskal–Wallis test). C Control and hsa_circ_0007813 knockdown cells were subjected to western blot analysis with anti-P-AMPK, anti-LC3, anti-P-S6K, anti-P-AKT, anti-P-ERK, anti-PARP1, anti-Vimentin, and anti-GAPDH. GAPDH was used as loading controls. D Kaplan–Meier plot of bladder cancer patients’ overall survival, grouped by median IGF2R expression level (Carried out by GEPIA). E Scatter plot presenting correlated expression levels between hsa_circ_0007813 and hsa-miR-361-3p in bladder cancer tissues (N = 30, generalized least squares and F-test). F Scatter plot presenting correlated expression levels between hsa_circ_0007813 and IGF2R in bladder cancer tissues (N = 30, generalized least squares and F-test). G Scatter plot presenting correlated expression levels between hsa-miR-361-3p and IGF2R (N = 30, generalized least squares and F-test). H Potential binding sites of hsa_circ_0007813/hsa-miR-361-3p and IGF2R mRNA 3′-UTR/hsa-miR-361-3p were located by Interactome. The third and sixth rows showed mutated hsa_circ_0007813 (MUT) and IGF2R mRNA 3′-UTR (MUT) sequence for dual-luciferase reporter assay. I HEK-293 cells were transfected with dual-luciferase reporter vectors carrying wild-type (WT-Circ) or mutated (MUT-Circ) hsa_circ_0007813 fragments. Cells were then transfected with control miRNA mimic (MimicCtrl) or hsa-miR-361-3p mimic (MimicMIR). At 48 h after transfection, luciferase activities were measured (N = 3, Kruskal–Wallis test). J HEK-293 cells were transfected with dual-luciferase reporter vectors carrying wildtype (WT-IGF2R) or mutated (MUT-IGF2R) IGF2R mRNA 3’-UTR fragments. Cells were then transfected with control miRNA mimic (MimicCtrl) or hsa-miR-361-3p mimic (MimicMIR). At 48 h after transfection, luciferase activities were measured (N = 3, Kruskal–Wallis test). All error bars in the figures indicated the standard deviation of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.

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